Yeast-generated virus-like particles as antigens for detection of human bocavirus 1-4 specific antibodies in human serum

Appl Microbiol Biotechnol. 2016 Jun;100(11):4935-46. doi: 10.1007/s00253-016-7336-8. Epub 2016 Feb 4.

Abstract

Human bocaviruses (HBoV) are non-enveloped, single-stranded DNA viruses, classified into the genus Bocavirus in the family Parvoviridae. Self-assembled virus-like particles (VLPs) composed of the major capsid protein VP2 of HBoV1-4 and mosaic VLPs composed of both VP2 and VP1 capsid proteins of HBoV1 were generated in yeast Saccharomyces cerevisiae and used to detect HBoV-specific IgG in human serum. Recombinant HBoV VLPs were similar to native HBoV particles in size and morphology. The prevalence of HBoV infection in a group of Lithuanian patients with clinical symptoms of respiratory tract infection was studied using purified yeast-generated VLPs as antigens in a competitive enzyme immunoassay (EIA). After depletion of cross-reactive antibodies, the seroprevalence of HBoV1 was 44.2 % and the seroprevalence of HBoV2-4 was 35.7 %. Mosaic VLPs consisting of HBoV1 VP1 and VP2 proteins showed a stronger reactivity with HBoV1 IgG-positive human serum specimens, and two equivocal serum specimens were reinterpreted as positive. Thus, mosaic VLPs offer a more sensitive tool for HBoV1 serology than currently available serodiagnostics tests based on VP2 VLPs. In conclusion, yeast S. cerevisiae represents an efficient expression system for generating recombinant HBoV1-4 VLPs of diagnostic relevance.

Keywords: ELISA; Human bocaviruses; Seroepidemiology; Virus-like particles; Yeast.

MeSH terms

  • Adolescent
  • Adult
  • Aged
  • Animals
  • Antibodies, Viral / blood*
  • Antigens, Viral / immunology*
  • Capsid Proteins / immunology
  • Child
  • Child, Preschool
  • Cloning, Molecular
  • Cross Reactions
  • Electrophoresis, Polyacrylamide Gel
  • Escherichia coli / genetics
  • Escherichia coli / metabolism
  • Female
  • Human bocavirus / isolation & purification*
  • Humans
  • Immunoglobulin G / blood
  • Infant
  • Lithuania
  • Male
  • Mice
  • Mice, Inbred BALB C
  • Middle Aged
  • Parvoviridae Infections / blood
  • Parvoviridae Infections / diagnosis*
  • Parvoviridae Infections / immunology
  • Recombinant Proteins / immunology
  • Recombinant Proteins / metabolism
  • Saccharomyces cerevisiae / metabolism
  • Specimen Handling
  • Young Adult

Substances

  • Antibodies, Viral
  • Antigens, Viral
  • Capsid Proteins
  • Immunoglobulin G
  • Recombinant Proteins